The chief objective of the first aspect of this investigation is to describe in detail the mechanism of action of flavoprotein oxidases and flavoprotein hydroxylases. Several lines of experimentation will be pursued to this end. Active site mapping of putrescine oxidase by the use of substrate analogs and site specific enzyme inactivators will be continued, and a full kinetic analysis of the enzyme will be concluded. Stopped-flow and steady state kinetic experiments will be used to test specific, proposed mechanisms. Further kinetic studies will be directed to lysine monooxygenase, a flavo-protein hydroxylase; subsequent work will then be directed toward other aspects of the mechanism of action of this hydroxylating enzyme. Effort will also be directed to the final development and exploitation of a unique rapid-scan spectrometer system for use with the stopped-flow spectrophotometer and on-line computer systems. In addition, construction of a unique circular dichroism (CD) spectrophotometer will be initiated. The on-line computer systems will also be equipped with a low cost magnetic disc data recording system to explore the possible advantages of this mode of data storage in the laboratory environment.